蒙药塔布森-2中9种苯丙素类成分在Caco-2细胞中的吸收特性研究

Absorption Characteristics of Nine Phenylpropanoids in Mongolian Medicine Tabson-2 Decoction in Caco-2 Cells

关键词:  塔布森-2  苯丙素类  Caco-2细胞  UPLC-MS/MS  P-糖蛋白

Key words:  Tabson-2 decoction  phenylpropanoids  Caco-2 cells  UPLC-MS/MS  P-glycoprotein

摘要:

目的 研究蒙药塔布森-2(Tabson-2 decoction，TBD)中苯丙素类成分在Caco-2细胞中的吸收特性，初步阐明TBD的口服吸收机制。方法 采用Caco-2细胞单层细胞模型，通过UPLC-MS/MS分析TBD在Caco-2细胞中的摄取成分，并建立UPLC-MS/MS分析方法测定TBD中吸收最好的9种苯丙素类成分原儿茶酸、新绿原酸、绿原酸、隐绿原酸、1,5-二咖啡酰奎宁酸、异绿原酸C、咖啡酸、二氢咖啡酸、3-羟基肉桂酸在Caco-2细胞中的含量，分别考察时间、浓度和P糖蛋白抑制剂对各成分的吸收影响。结果 研究发现咖啡酸、二氢咖啡酸的总摄取量在0~180 min为上升趋势，未呈现饱和；3-羟基肉桂酸在90 min左右吸收恒定趋于饱和；隐绿原酸、1,5-二咖啡酰奎宁酸、异绿原酸C、新绿原酸、绿原酸、原儿茶酸的摄取量约从90 min开始，随时间的增加先下降后上升。与苯丙素类成分相比，加入P-糖蛋白抑制剂维拉帕米及环孢菌素A后对二氢咖啡酸的吸收产生了影响，说明二氢咖啡酸是P-糖蛋白底物。结论 TBD主要苯丙素类成分进入Caco-2细胞以被动扩散为主，主动转运为辅，且除二氢咖啡酸外其他8个成分的吸收过程不受P-糖蛋白的外排作用影响。

Abstract:

OBJECTIVE To study the absorption characteristics of phenylpropanoids of Mongolian medicine Tabson-2 decoction(TBD) in Caco-2 cells and to preliminarily clarify the oral absorption mechanism of TBD. METHODS Caco-2 cell monolayer model was used to analyze the uptake components of TBD in Caco-2 cells by UPLC-MS/MS, and UPLC-MS/MS analysis method was established to determine the nine best absorbed components of TBD, protocatechuic acid, neochlorogenic acid, chlorogenic acid, cryptogenic acid, 1,5-dicaffeinate quinic acid, isochlorogenic acid C, caffeic acid, dihydrocaffeic acid, chlorogenic acid. The effects of time, concentration and P-glycoprotein inhibitor on the absorption of each component were investigated. RESULTS The overall intake of caffeic acid and dihydrocaffeic acid showed an upward trend in 0-180 min, and did not show saturation. The absorption of 3-hydroxycinnamic acid was constant at about 90 min and tended to saturation. The intakes of cryptochlorogenic acid, 1,5-dicaffeinate, quinic acid, isochlorogenic acid C, neochlorogenic acid, chlorogenic acid and protocatechuic acid first decreased and then increased with time from about 90 min. The addition of P-glycoprotein inhibitor verapamil and cyclosporin A had an effect on the absorption of dihydrocaffeic acid compared with the phenylpropanoid components, indicated that dihydrocaffeic acid was the substrate of P-glycoprotein. CONCLUSION The main phenylpropanoids of TBD enter Caco-2 mainly by passive diffusion, supplemented by active transport, and the absorption process of the other eight components is not affected by the efflux of P-glycoprotein except dihydrocaffeic acid.